Microbiological stabilization of tiger nuts' milk beverage using ultra-high pressure homogenization. A preliminary study on microbial shelf-life extension.

Tiger nuts' milk beverages are highly perishable products. For this reason, the interest of food industry for their commercialization makes necessary the application of preservation treatments to prolong their shelf-life. In the current study, the effect of ultra-high pressure homogenization (UHPH) on the microbiological and sensory qualities of tiger nuts' milk beverage was evaluated. Characteristics of UHPH-treated products (at 200 and 300 MPa, with inlet temperature of 40 °C) were compared with those of raw (RP) and conventionally homogenized-pasteurized (H-P) beverages, after treatment and during cold storage at 4 °C. Microbiological quality of beverages was studied by enumerating total counts, psychrotrophic bacteria, lactobacilli, enterobacteria, molds and yeasts, and mesophilic spores. Evolution of color and sensory characteristics of beverages were also determined. Microbiological shelf-life of the tiger nuts' milk beverages was extended from 3 to 25, 30 and 57 days by applying H-P and UHPH treatments at 200 and 300 MPa, respectively. Color of beverages was the only attribute that differentiated UHPH samples from the others, with greater luminosity and whiteness. Hence, UHPH treatments showed to be an alternative to the conventional H-P for obtaining tiger nuts' milk beverages with an improved microbiological shelf-life and good sensorial characteristics.

Single-trial conditioning in a human taste-endotoxin paradigm induces conditioned odor aversion but not cytokine responses.

Immunological responses to bacterial endotoxin can be behaviorally conditioned in rodents. However, it is unclear whether an acute systemic inflammatory response can be behaviorally conditioned in humans. Thus, in a double-blind placebo-controlled study, 20 healthy, male subjects received either a single injection of lipopolysaccharide (LPS) or saline together with a novel tasting beverage (conditioned stimulus, CS). Five days later, all subjects received a saline injection and were re-exposed to the CS. Blood was drawn prior to as well as 0.5, 1.5, 3, 4, 6, and 24 h after LPS administration or CS re-exposure. Endotoxin administration led to transient increases in plasma concentrations of interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α and to a significant rise in body temperature. Sole presentation of the CS during evocation did induce neither alterations in body temperature nor changes in plasma cytokine levels. However, subjects in the experimental group rated the smell of the CS significantly more aversive compared to the control group. Employing endotoxin as a US in a single trial taste-immune conditioning paradigm in humans shows a behaviorally conditioned smell aversion but no learned alterations in cytokine levels.

Mice do not develop conditioned taste aversion because of immunity loss.

OBJECTIVES: This study intends to test the generation of conditioned taste aversion and conditioned immunodepression by daily paired administration of saccharin solution with cyclophosphamide, 15 mg/kg, for 4 days. METHODS: One group of male mice of the outbred CD1 strain drank 0.15% saccharin and received 1 injection of cyclophosphamide, 15 mg/kg, for 4 days (paired group), another group (unpaired group) received the same doses of saccharin and cyclophosphamide noncontingently, the third group (cy60) received saccharin paired with cyclophosphamide, 60 mg/kg, and the fourth group (placebo) received saccharin in the absence of cyclophosphamide. All mice were immunized with keyhole limpet hemocyanin (KLH), 0.2 mg, 1 day before the treatments. RESULTS: Mice of the paired, unpaired and cy60 groups displayed a similarly decreased antibody response to KLH, but mice of the paired group did not develop an aversion to saccharin while mice of the cy60 group did. Besides, repeat presentation of saccharin to mice of the paired group did not alter their antibody response to ovalbumin compared with mice of the unpaired or placebo group. CONCLUSIONS: Taste aversion was not elicited in response to impaired immunity and the conditioned stimulus (saccharin) did not impair the antibody response.

To Detect and Reject, Parallel Roles for Taste and Immunity.

PURPOSE OF REVIEW: From single cells to entire organisms, biological entities are in constant communication with their surroundings, deciding what to 'allow' in, and what to reject. In very different ways, the immune and taste systems both fulfill this function, with growing evidence suggesting a relationship between the two, through shared signaling pathways, receptors, and feedback loops. The purpose of this review was to explore recent reports on taste and immunity in model animals and in humans to explore our understanding of the interplay between these systems. RECENT FINDINGS: Acute infections in the upper airway, as with SARS-CoV-2, are associated with a proinflammatory state, and blunted taste perception. Further, recent findings highlight taste receptors working as immune sentinels throughout the body. Work in humans and mice also points to inflammation from obesity impacting taste, altering taste bud abundance and composition. There is accumulating evidence that taste cells, and particularly their receptors, play a role in airway and gut immunity, responsive to invading organisms. Inflammation itself may further act on taste buds and other taste receptor expressing cells throughout the body as a form of homeostatic control.

The conditioned stimulus elicits taste aversion but not sickness behavior in conditioned mice.

OBJECTIVES: This article extends previous reports on (i) elicitation of taste aversion after pairing a flavored beverage (saccharin solution) with a disease-provoking microbial product (lipopolysaccharide, LPS, or polyinosinic:polycytidylic acid, poly I:C); (ii) elicitation of sickness behavior (assessed as diminished ingestion of water and food) by the conditioned stimulus, and (iii) development of tolerance to those microbial products. METHODS: Mice of the CD1 strain were conditioned by pairing ingestion of 0.15% saccharin solution with injection of LPS (100 mug/mouse) or poly I:C (6 mg/kg). A few days later, some mice were offered saccharin solution and were injected with saline, whereas other mice were offered saccharin solution and were injected with the microbial product. RESULTS: Regardless of the nature of the unconditioned stimulus (LPS or poly I:C), (i) taste aversion to saccharin ensued, (ii) tolerance ensued to sickness elicitation by a second administration of the microbial component, and (iii) saccharin taste did not evoke sickness. CONCLUSIONS: Symptoms of infectious sickness in the absence of infection are hardly explained by exposure to the conditioned stimulus.

Monosodium glutamate 'allergy': menace or myth?

Monosodium glutamate (MSG) is a salt form of a non-essential amino acid commonly used as a food additive for its unique flavour enhancing qualities. Since the first description of the 'Monosodium glutamate symptom complex', originally described in 1968 as the 'Chinese restaurant syndrome', a number of anecdotal reports and small clinical studies of variable quality have attributed a variety of symptoms to the dietary ingestion of MSG. Descriptions of MSG-induced asthma, urticaria, angio-oedema, and rhinitis have prompted some to suggest that MSG should be an aetiologic consideration in patients presenting with these conditions. This review prevents a critical review of the available literature related to the possible role of MSG in the so-called 'Chinese restaurant syndrome' and in eliciting asthmatic bronchospasm, urticaria, angio-oedema, and rhinitis. Despite concerns raised by early reports, decades of research have failed to demonstrate a clear and consistent relationship between MSG ingestion and the development of these conditions.

Immune cells of the human peripheral taste system: dominant dendritic cells and CD4 T cells.

Taste loss or alterations can seriously impact health and quality of life due to the resulting negative influence on eating habits and nutrition. Infection and inflammation are thought to be some of the most common causes of taste perception disorders. Supporting this view, neuro-immune interactions in the peripheral gustatory system have been identified, underlying the importance of this tissue in mucosal immunity, but we have little understanding of how these interactions influence taste perception directly or indirectly. This limited understanding is evident by the lack of even a basic knowledge of the resident immune cell populations in or near taste tissues. The present study characterized the distribution and population of the major immune cells and their subsets in healthy human anterior, lingual, fungiform papillae (FP) using immunohistochemistry. Dendritic cells (DCs) were the predominant innate immune cells in this tissue, including four subtypes: CD11c(+) DCs, DC-SIGN+ immature DCs, CD83(+) mature DCs, and CD1a(+) DCs (Langerhans cells). While most DCs were localized beneath the lamina propria and only moderately in the epithelium, CD1a(+) Langerhans cells were exclusively present within the epithelium and not in sub-strata. A small number of macrophages were observed. T lymphocytes were present throughout the FP with CD4(+) T cells more prevalent than CD8(+) T cells. Very few CD19(+) B lymphocytes were detected. The results show that DCs, macrophages, and T lymphocytes are the constitutive guardians of human FP taste tissue, with DCs and CD4 T cells being dominant, while B lymphocytes are rare under normal, healthy conditions. These observations provide a basic anatomical foundation for the immune response in the healthy human tongue as a basis for subsequent disease-related studies, but none of the present data indicate that the immune cell populations identified are, in fact, altered in individuals with abnormal taste perception.

Lipopolysaccharide dose dependently impairs rapid toxin (LiCl)-induced gustatory conditioning: a taste reactivity examination of the conditioned taste aversion.

There is much debate on how immune activation affects cognitive processing. Research has shown that stimulation of the immune system can significantly impair, have no adverse effects, or enhance learning and memory processes in animals. The present experiment evaluated the effects of the bacterial endotoxin, lipopolysaccharide (LPS) on the acquisition of a rapidly acquired conditioned taste aversion using a toxin-containing food. Male Long Evans rats were fitted with intraoral cannulae and habituated to the taste reactivity procedure. Rats received two conditioning days, 72 h apart, in which they were injected systemically with LPS (200, 100, or 50 microg/kg) or NaCl (0.9% vehicle) and 90 min later placed in the taste reactivity test chamber. Rats were given 5 brief (1 min) intraoral infusions of either a LiCl-adulterated sucrose solution (0.15M LiCl+0.3M sucrose) or NaCl-sucrose solution (0.15M NaCl+0.3M sucrose) across a 1h period. On the test day (72 h after the last conditioning trial), rats were given a 2 min intraoral infusion of the respective taste in a drug-free state. Individual taste reactivity responses were recorded and analyzed. Results demonstrate that rats treated with LPS dose-dependently increased ingestive responding to the LiCl-sucrose flavor while at the same time showing reduced rejection response frequency on the two conditioning days. LPS treatment did not alter taste reactivity responding to the NaCl-sucrose solution. On the test day, the LPS groups again displayed a dose dependent increase in ingestive responses and a decrease in rejection responses to the LiCl-sucrose taste. The present results suggest that LPS-induced immune system activation, significantly impairs the rapid acquisition of a conditioned taste aversion.

Taste-aversion conditioning, but not immunosuppression conditioning, occurs under partial water deprivation.

The authors investigated whether conditioned taste aversion and immunosuppression took place when water was available during conditioning and test protocols. The authors elicited taste-aversion conditioning and immunosuppression in outbred CD1-strain mice by pairing a conditioned stimulus (sucrose or saccharin solution) with an unconditioned stimulus (cyclophosphamide) that causes gastrointestinal upset and is immunosuppressive. The authors introduced a new conditioning protocol: 5 pairings of a saccharin solution with a low-dose injection of cyclophosphamide. Under these conditions, the authors generated conditioned aversion to saccharin but did not generate conditioned decrease of the antibody response. The authors conclude that taste-aversion conditioning, but not immunosuppression conditioning, occurred under partial water deprivation.

Weakened [corrected] taste-LPS association during endotoxin tolerance.

In naive individuals, the administration of bacterial lipopolysaccharide (LPS) provokes a rapid systemic increase in pro-inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6, inducing an acute phase response including sickness behavior. Strong associative learning occurs when relevant gustatory/olfactory stimuli precede the activation of the immune system, affecting long-term individual food selection and nutritional strategies. Repeated LPS administration results in the development of an endotoxin tolerance status, characterized by a drastic reduction in the LPS-induced cytokine response. Here we investigated how the postprandial categorization of a relevant taste (0.2% saccharin) changed after administration of a high dose of LPS (0.5 mg/kg i.p.) in LPS-tolerant animals. Determination of the consummatory fluid intake revealed that, in contrast to LPS-naive rats, taste-LPS association did not occur during endotoxin tolerance. Ninety minutes after the single association trial, the plasma responses of TNF-alpha, IL-1beta and IL-6 were completely blunted in LPS-tolerant animals, which also resulted in low LPS-adipsogenic and LPS-anorexic effects. These findings indicate that an identical immune challenge can result in completely different neuro-behavioral consequences depending on the immune history of the individual, thus revealing part of the complex interconnection between the immune and neuro-endocrine systems in regulating food selection and consumption during the infectious process.

Characterization of the therapeutic properties of Chinese herbal materials by measuring delayed luminescence and dendritic cell-based immunomodulatory response.

Based on the traditional Chinese medicine theory, the Chinese pharmacopeia assigns a therapeutic description of "taste" to all herbs; thus, an herb's "taste" is valued in traditional Chinese medicine as a major ethnopharmacological category and reflects the herb's therapeutic properties. These properties guide the practitioner with respect to preparing a specific herbal formula in order to provide each patient with a personalized intervention. The key challenge in evidence-based medicine is to characterize herbal therapeutic properties from a multi-target, multi-dimensional systems pharmacology perspective. Here, we used delayed luminescence (DL, the slowly decaying emission of photons following excitation with light) as a rapid, direct, highly sensitive indicator to characterize the properties of herbal medicines. The DL parameters were able to reliably identify a specific category of herbal materials with the so-called "sweet" taste. To support the DL results and provide biological relevance to the DL results, we used a murine bone marrow-derived dendritic cell-based assay to examine the immunomodulatory effects of herbal extracts from various "taste" categories. Our results indicate that DL may serve as a robust and sensitive tool for evaluating the therapeutic properties of herbs based on the traditional Chinese medicine classification of "taste". Thus, DL provides a promising technological platform for investigating the properties of Chinese herbal medicines both qualitatively and quantitatively.

Up-regulation of activated macrophages in response to degeneration in the taste system: effects of dietary sodium restriction.

Dietary sodium restriction combined with unilateral chorda tympani nerve section leads to a rapid and specific decrease in neurophysiological taste responses to sodium in the contralateral, intact chorda tympani (Hill and Phillips [1994] J. Neurosci. 14:2904-2910). Previous work demonstrated that dietary sodium restriction may induce these early functional deficits by inhibiting immune activity after denervation (Phillips and Hill [1996] Am. J. Physiol. 271:R857-R862). However, little is known about the leukocyte response to denervation of taste buds in fungiform papillae. In the current study, it was hypothesized that T cells and macrophages are increased in the tongue after unilateral denervation in control-fed but not sodium-restricted animals. Adult, specified pathogen-free rats received unilateral chorda tympani nerve section or sham section followed by dietary sodium restriction or maintenance on control diet. At day 1, 2, 5, 7, or 50 postsectioning, immunostaining was used to detect the percentage of staining for activated macrophages, the number of alpha beta T cells, and the number of delta gamma epithelial T cells in the tongue. The number of lingual T cells did not significantly differ between treatment groups following denervation. However, there was a dramatic bilateral increase in ED1(+) staining for activated macrophages in control-fed rats that peaked at day 2 postsectioning. In contrast, sodium-restricted rats did not show an increase in activated macrophages above baseline at any time postsectioning. Further analysis of extralingual macrophages indicated that the deficit in immune activity in sodium-restricted rats is localized to the tongue and is not widespread. A model for immune modulation of taste receptor cell function is proposed based on these novel findings.

Immune activation paired with intraoral sucrose conditions oral rejection.

The effects of lipopolysaccharide (LPS) and LiCl on conditioned taste aversion acquisition using intraoral infusions as the method of taste delivery was examined. Rats received two pairings of an intraorally delivered sucrose (5 ml) taste with the effects of a systemic injection of LPS, LiCl or NaCl. The magnitude of conditioning was quantified by scoring taste reactivity responses to a brief intraoral infusion of sucrose in the absence of any drug injection. Rats previously conditioned with LiCl or LPS displayed clear evidence of conditioned aversion with increased oral rejection responses relative to saline controls. Our results suggest activation of the immune system with LPS can condition consummatory aspects of ingestion when this conditioning involves intraoral fluid presentation.

Topographic organization of Fos-like immunoreactivity in the rostral nucleus of the solitary tract evoked by gustatory stimulation with sucrose and quinine.

Fos immunohistochemistry was used to elucidate the pattern of activation elicited by two qualitatively and hedonically distinct taste stimuli, sucrose and quinine, within the first-order gustatory relay, the rostral division of the nucleus of the solitary tract. Compared to unstimulated controls, both sucrose and quinine elicited significant increases in Fos-like immunoreactivity in the rostral central subnucleus, the region of the rostral solitary nucleus that receives the densest primary afferent input. Within the rostral central subnucleus, neurons that exhibited Fos-like immunoreactivity following quinine stimulation were concentrated medially, but neurons that exhibited Fos-like immunoreactivity following sucrose stimulation were distributed more evenly along the mediolateral axis. Despite their differential distribution, sucrose- and quinine-activated neurons also demonstrated notable intermingling. Further, the chemotopic arrangement was only partially consistent with what would be predicted if chemotopy was merely an outcome of orotopy. Our results suggest that a rough chemotopy characterizes the organization of taste responses in the nucleus of the solitary tract, and that the topographic pattern of taste afferent terminations in this nucleus is related to their chemosensitivity as well as to their peripheral spatial distribution.

Conditioned taste aversion produced by cyclosporine A: concomitant reduction in lymphoid organ weight and splenocyte proliferation.

The classical conditioning of immune parameters is commonly conducted within a conditioned taste aversion (CTA) paradigm. In this study, the immunosuppressive drug cyclosporine A (CsA) was investigated for its ability to produce both taste aversion to a novel stimulus and conditioned alterations in immune functioning. The paradigm comprised the pairing of a 0.2% saccharin solution (the conditioned stimulus; CS) with an intraperitoneal injection of 20 mg/kg CsA (the unconditioned stimulus; UCS). Upon saccharin re-presentation, a marked reduction in fluid consumption was observed, indicating aversion to the novel substance (=CTA). By using a single CsA/saccharin pairing the CTA lasted for one CS representation. However, by implementing three pairings, this effect could be extended for up to seven representations. No noticeable difference was recorded by adjusting the saccharin representation from every consecutive day to every second day. The most effective paradigm in creating CTA was subsequently investigated for its effectiveness in producing conditioned immune alterations. Animals were killed on the day of the third CS re-presentation, and immune functions assessed. Conditioned animals displayed a significant reduction in thymus and spleen weights. Effects on the spleen were further investigated, revealing a significantly reduced proliferative ability of isolated splenocytes to concanavalin A. These results demonstrate that the physiological effects produced by CsA are sufficiently salient to elicit CTA. Furthermore, the reduction in lymphoid organ weight and splenocyte proliferation induced by CsA are also conditionable using this paradigm.

Immunological induction of flavor aversion in mice.

1. Young adult BALB/c and B6D2F1 mice of both sexes (20 +/- 2 g) immunized ip with 2 doses of 10 micrograms ovalbumin (Ova), but not with 2 doses of 10 micrograms bovine gammaglobulins (BGG), show aversion to the ingestion of sweetened egg white or crystallized Ova solutions which are avidly ingested by normal mice. In 24 h, normal mice or mice immunized with BGG ingested, respectively, 340 +/- 80 and 265 +/- 56 mg of sweetened egg white per gram of body weight (mg/gbw); in the same period, Ova-immunized mice ingested less than one tenth these amounts (18 +/- 5 mg/gbw). ELISA-titers of anti-Ova and anti-BGG antibodies in immune mice were of similar magnitude. 2. Aversion arises coincidentally with the emergence of anti-ovalbumin antibodies in serum in the primary response, 14 days after primary immunization. 3. Previous induction of oral tolerance to ovalbumin by a single gavage with 20 mg Ova 7 days before primary ip immunization, which blocks the increase of specific antibodies in serum, also blocks the development of the aversive phenomenon. 4. Aversion was induced to 1 mg/ml but not 0.1 mg/ml sweetened crystallized ovalbumin solutions and was already noticeable 2 h after exposure of immunized mice to sweetened egg white solutions. 5. We conclude that, at least in experimental situations, immunological factors may be of decisive importance in diet selection.

Influenza vaccinations and chemosensory function.

BACKGROUND: Although influenza vaccines have saved millions of lives, some have been associated with extremely rare adverse effects such as Guillain-Barré syndrome, Bell's palsy, and optic neuritis. Despite the fact that olfactory loss after an influenza vaccination is noted in one case report, no quantitative olfactory testing was performed. Hence, it is unclear whether, in fact, olfactory dysfunction can be associated with such vaccinations. This study was designed to (1) identify patients from the University of Pennsylvania Smell and Taste Center who attributed their empirically determined chemosensory disturbances to influenza vaccinations and (2) determine whether influenza vaccinations add to the degree of olfactory or gustatory dysfunction due to other causes. METHODS: A retrospective analysis of self-reported etiologies of 4554 consecutive patients presenting to the University of Pennsylvania Smell and Taste Center with complaints of chemosensory dysfunction was performed. Those who reported dysfunction secondary to influenza vaccinations were identified. Additionally, in a subset of 925 patients for whom detailed inoculation histories were available, it was determined whether the number of lifetime inoculations added to the deficits due to other causes. RESULTS: Nine of the 4554 patients (0.19%) attributed olfactory disturbances to an influenza vaccination. None complained of taste dysfunction. All nine had abnormally low scores on the University of Pennsylvania Smell Identification Test (p < 0.001), with three being anosmic and six microsmic. Seven had elevated phenyl ethyl alcohol detection thresholds (p < 0.05). Two cases exhibited mild-to-moderate loss of whole mouth taste function. Of the 925 patients, no association was evident between the number of lifetime vaccinations and the chemosensory test scores. In accord with previous studies, age and sex were significantly related to the test scores. CONCLUSION: A very small percentage of the 4554 patients evaluated (0.19%) attributed their chemosensory dysfunction to a prior influenza vaccination. No influences of the number of lifetime influenza vaccinations on the test scores were evident in the subset of 925 patients whose dysfunction was due to other causes.

Learned immunosuppression: extinction, renewal, and the challenge of reconsolidation.

Behavioral conditioning of immune responses is one of the most impressive examples for the bidirectional communication among the nervous and immune systems. We established a model of behaviorally conditioned immunosuppression employing a conditioned taste aversion (CTA) paradigm in the rat pairing a novel taste (saccharin) as a conditioned stimulus (CS) with the immunosuppressive drug cyclosporine A (CsA) as an unconditioned stimulus (US). By re-presenting the CS during evocation, rats avoid drinking the saccharin. Concomitantly animals display an immunosuppression reflected by an ex vivo reduction in splenic T cell proliferation as well as diminished interleukin-2 and interferon-γ production and cytokine mRNA expression, mimicking the actual effect of the US (CsA). Due to the fact that the kinetics of this behaviorally conditioned immunosuppression are completely unknown, extinction of the conditioned response on the behavioral level (CTA) as well as in the immune response needs to be elucidated together with the neural processes mediating the extinction process.